Biallelic <i>β‐carotene oxygenase 2</i> knockout results in yellow fat in sheep via <scp>CRISPR</scp>/Cas9

Niu, Yiyuan; Jin, Min Kyung; Li, Y.; Li, Pengmin; Zhou, Jianghua; Wang, X.; Petersen, Björn; Huang, Xingxu; Kou, Qifang; Chen, Y.

doi:10.1111/age.12515

Cited by 33 publications

(24 citation statements)

References 12 publications

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“…When SCNT is mastered by the laboratory, the low efficient rate will be compensated with most of the healthy born animals carrying the desired mutation. Genome editing mediated by SCNT is applied by some laboratories in some kind of projects (e.g., multiplex editing), but the high efficiency of CRISPR after direct microinjection into zygotes has allowed an easier approach (sheep: [6,7,15,16]; goats: [9,17]; pigs: [11,13]). All together, these reports showed that with CRISPR it is possible to perform direct microinjection into the zygote of different species, with minimum effects on developmental competence and pregnancy rates, high newborn survival rate, and high editing rate with acceptable homozygous proportion.…”

Section: Embryo Manipulationmentioning

confidence: 99%

“…These often disrupt open reading frames, effectively creating gene knockouts. This mechanism led to the generation of the first successful disruption of endogenous genes by CRISPR-Cas system in livestock (sheep [6,7], goats [8,9], cows [10] and pigs [11e14]). On the other hand, the HDR pathway employs a homologous repair template to fix the DSB.…”

Section: Introductionmentioning

confidence: 99%

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CRISPR in livestock: From editing to printing

et al. 2020

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a b s t r a c tPrecise genome editing of large animals applied to livestock and biomedicine is nowadays possible since the CRISPR revolution. This review summarizes the latest advances and the main technical issues that determine the success of this technology. The pathway from editing to printing, from engineering the genome to achieving the desired animals, does not always imply an easy, fast and safe journey. When applied in large animals, CRISPR involves time-and cost-consuming projects, and it is mandatory not only to choose the best approach for genome editing, but also for embryo production, zygote microinjection or electroporation, cryopreservation and embryo transfer. The main technical refinements and most frequent questions to improve this disruptive biotechnology in large animals are presented. In addition, we discuss some CRISPR applications to enhance livestock production in the context of a growing global demand of food, in terms of increasing efficiency, reducing the impact of farming on the environment, enhancing pest control, animal welfare and health. The challenge is no longer technical. Controversies and consensus, opportunities and threats, benefits and risks, ethics and science should be reconsidered to enter into the CRISPR era.

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Section: Embryo Manipulationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

CRISPR in livestock: From editing to printing

et al. 2020

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“…Seven ASE SNPs were identified in BCO2 gene from the samples of subcutaneous fat and tail fat, six of these ASE SNPs were located in exon region of the gene and the remaining one ASE SNP was located in its 3'-UTR region. Knocking out the biallelic BCO2 gene could lead to yellow fat in Tan sheep and a nonsense mutation in this gene is strongly related to the yellow fat phenotype in Norwegian sheep [76,77]. A total of seven ASE SNPs were found in PM20D1 gene, which involved in the cellular lipid process.…”

Section: Discussionmentioning

confidence: 99%

Allele-specific expression reveals the phenotypic differences between thin- and fat-tailed sheep

Shao¹,

He²,

Pan³

et al. 2020

Preprint

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Background: The thin-tailed sheep breeds from Europe and the fat-tailed sheep breeds from China exhibit distinct phenotypic differences in fat deposition and meat production traits. However, the molecular mechanisms underlying gene expression related to these phenotypic differences are not well understood. Allele-specific expression (ASE) refers to the significant imbalance of expression levels of two parental alleles. Characterization of such events in F1 hybrid offspring generated from these two groups of sheep breeds can minimize the external factors influencing gene expression and reveal the variants with a cis -regulatory effect on gene expression. The aim of the present study was to investigate the genetic factors that influence different fat-deposition and meat production traits between thin- and fat-tailed sheep.Results: Fifteen F1 hybrids were generated from crosses between Texel and Kazakh sheep as the representative phenotypes of thin- and fat-tailed breeds, respectively. Totally, 33 whole genomes from F1 individuals and their parents were sequenced with an average depth of ~17.21× coverage per sample. ASE analysis results from 70 RNA-seq samples of adipose and skeleton muscle tissues showed 128 ASE candidate genes were related to the function of fat deposition and meat production traits. A genome-wide scan of selective sweeps was also conducted between these two groups of sheep breeds in an effort to identify genomic regions related to fat deposition and meat production, respectively. We detected signatures of selection in ASE genes associated with fat deposition (e.g., PDGFD ) and meat production traits (e.g., LRCC2 ). Further analysis suggested that PDGFD and LRCC2 genes were speculated to be causative genes for fat deposition and meat production traits in sheep, respectively. Furthermore, AMPK signaling pathway was significantly enriched in ASE genes related to fatty acid biosynthesis in both adipose and skeleton muscle tissues, while PPAR signaling pathway was significantly enriched in ASE genes related to lipid metabolism in adipose tissue. Conclusions: Our finding illustrates that the expression of identified ASE genes could potentially lead to the differences in traits of fat deposition and meat production between thin- and fat-tailed sheep. Keywords: allele-specific expression, phenotypic difference, thin- and fat-tailed sheep, whole-genome sequencing, transcriptome

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“…Another transgenic pig exhibited an increased content of Omega‐3 fatty acids in meat due to the expression of the fatty acid desaturase fat‐1 from Caenorhadbitis elegans (Lai et al., ). Using CRISPR, an increase in yellow fat in sheep meat has been recently achieved by removing beta‐carotene oxygenase 2 (BCO2) (Niu et al., ).…”

Section: Improved Animal Productsmentioning

confidence: 99%

CRISPR is knocking on barn door

Lamas-Toranzo

Guerrero-Sánchez

Miralles-Bover

et al. 2017

Reprod Domestic Animals

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Genome modification at specific loci in livestock species was only achievable by performing homologous recombination in somatic cells followed by somatic cell nuclear transfer. The difficulty and inefficiency of this method have slowed down the multiple applications of genome modification in farm animals. The discovery of site-specific endonucleases has provided a different and more direct route for targeted mutagenesis, as these enzymes allow the ablation (KO) or insertion (KI) of specific genomic sequences on a single step, directly applied to zygotes. Clustered regularly interspaced short palindromic repeats (CRISPR), the last site-specific endonuclease to be developed, is a RNA-guided endonuclease, easy to engineer and direct to a given target site. This technology has been successfully applied to rabbits, swine, goats, sheep and cattle, situating genome editing in livestock species at an attainable distance, thereby empowering scientist to develop a myriad of applications. Genetically modified livestock animals can be used as biomodels to study human or livestock physiology and disease, as bioreactors to produce complex proteins, or as organ donors for transplantation. Specifically on livestock production, genome editing in farm animals may serve to improve productive genetic traits, to improve various animal products, to confer resistance to diseases or to minimize the environmental impact on farming. In this review, we provide an overview of the current methods for site-specific genome modification in livestock species, discuss potential and already developed applications of genome edition in farm animals and debate about the possibilities for approval of products derived from gene-edited animals for human consumption.

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Biallelic β‐carotene oxygenase 2 knockout results in yellow fat in sheep via CRISPR/Cas9

Cited by 33 publications

References 12 publications

CRISPR in livestock: From editing to printing

CRISPR in livestock: From editing to printing

Allele-specific expression reveals the phenotypic differences between thin- and fat-tailed sheep

CRISPR is knocking on barn door

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